Anti dr5

Manufactured by R&D Systems

Sourced in United States

Anti-DR5 is a recombinant human protein that binds to the Death Receptor 5 (DR5) also known as TRAIL-R2. DR5 is a member of the tumor necrosis factor receptor superfamily and plays a role in the induction of apoptosis.

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4 protocols using anti dr5

TRAIL-induced Apoptosis Regulation Mechanisms

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Cannabidiol and VAS2870 were purchased from Sigma (St. Louis, MO, USA). TRAIL and anti-DR5 were purchased from R&D Systems (Minneapolis, MN, USA). Anti-Bak, anti-Bcl-2, anti-Mcl-1, anti-Bcl-xL, and anti-DR4 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-XIAP, anti-NOXA, anti-BIM, anti-survivin, anti-Bid, anti-IRE1α, anti-phospho-IRE1α, anti-Bip, anti-GRP94, anti-ATF6, anti-eIF2α, anti-phospho-eIF2α, anti-CHOP, anti-cleaved PARP, anti-caspase-3, and anti-caspase-9 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). The anti-actin antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA). For the secondary antibodies, anti-mouse IgG horseradish peroxidase (HRP) and anti-rabbit IgG HRP were purchased from Cell Signaling Technology.

Kim J.L., Kim B.R., Kim D.Y., Jeong Y.A., Jeong S., Na Y.J., Park S.H., Yun H.K., Jo M.J., Kim B.G., Kim H.D., Kim D.H., Oh S.C., Lee S.I, & Lee D.H. (2019). Cannabidiol Enhances the Therapeutic Effects of TRAIL by Upregulating DR5 in Colorectal Cancer. Cancers, 11(5), 642.

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Protein Expression Analysis by Western Blot

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Total protein concentrations of supernatant fractions were determined using the bicinchoninic acid (BCA) protein assay (Bio-Rad, Inc.). Equal amounts of protein aliquots were boiled in equal volumes of 2 × SDS Laemmli sample buffer and resolved by 8% or 10% (wt/vol) with primary antibodies: anti-TRAIL (0.2 µg/mL; R&D System Inc., Minneapolis, MN), anti-PDL-1 (0.1 µg/ml; Cell Signaling Inc.), anti-DR5 (1 µg/mL; R&D System Inc.), anti-mDcR1 (1 µg/mL; R&D System Inc.), and anti-β-actin (2 µg/ml; Abcam Inc., UK). β-actin was used as an internal loading control.

Ji Y.W., Lee J.H., Choi E.Y., Kang H.G., Seo K.Y., Song J.S., Kim H.C, & Lee H.K. (2020). HIF1α-mediated TRAIL Expression Regulates Lacrimal Gland Inflammation in Dry Eye Disease. Investigative Ophthalmology & Visual Science, 61(1), 3.

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Molecular mechanisms of TRAIL-induced apoptosis

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6-Gingerol was purchased from LKT Laboratories (St. Louis, MO, USA). Treatments of drugs were accomplished by aspirating the medium and replacing it with medium containing these drugs. Soluble human recombinant SuperKillerTRAIL (referred to as TRAIL in this manuscript) was purchased from Enzo Biochemicals (Enzo Life Sciences), diluted, and stored in KillerTRAIL storage and dilution buffer (Enzo Life Sciences). 6-Carboxy-2′,7′-dichlorofluorescein diacetate (H2DCF-DA) and dihydroethidium (DHE) were from Molecular Probe. N-acetylcysteine was from Sigma. Anti-c-FLIP, anti-Bax, anti-Bcl-2, and anti-p53 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-XIAP, anti-survivin, anti-Bid, anti-phospho ERK and anti-ERK, anti-phospho-p38, anti-p38, anti-cleaved caspase-3, anti-caspase-8, and anti-PARP-1 were purchased from Cell Signaling (Beverly, MA, USA). Anti-phospho JNK and anti-JNK were purchased from Upstate Biotechnology (Lake Placid, NY, USA). Anti-Flag M2 were purchased from Sigma (USA). Anti-DR4 and anti-DR5 were purchased from R&D Systems (Plymouth Metting, PA, USA). Anti-actin antibody was purchased from MP Biomedicals (Solon, OH, USA). For the secondary antibodies, anti-mouse-IgG-HRP and anti-rabbit-IgG-HRP were purchased from Santa Cruz Biotechnology.

Lee D.H., Kim D.W., Jung C.H., Lee Y.J, & Park D. (2014). Gingerol sensitizes TRAIL-induced apoptotic cell death of glioblastoma cells. Toxicology and applied pharmacology, 279(3), 253-265.

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Modulation of Autoimmune Inflammation

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Unless otherwise specified, reagents were injected into mice on days 1 and 3. A total of 1 μg of recombinant mouse sFasL, sTRAIL, or CX3CL1 (R and D Systems, Minneapolis, MN) was injected intraperitoneally and 50 μg of anti-mouse FasL, anti-Fas, anti-DR5, or anti-CX3CR1 antibodies (R and D Systems) was injected intravenously. To inhibit apoptosis in vivo, 50 μg of Z–VAD–FMK (Calbiochem, Billerica, MA,) was administered. The vehicle and isotype control injection were administered in the same way. To transfer splenocytes into Fasl^gld/gld mice, mouse spleens were homogenized and treated with red blood cell lysis solution (Qiagen, Hilden, Germany). In total, 1 × 10⁶ spleen cells were pooled in phosphate-buffered saline (PBS) and injected intravenously into Fasl^gld/gld mice on the day before K/BxN serum injections.

Jeong D., Kim H.S., Kim H.Y., Kang M.J., Jung H., Oh Y., Kim D., Koh J., Cho S.Y., Jeon Y.K., Lee E.B., Lee S.H., Shin E.C., Kim H.M., Yi E.C, & Chung D.H. (2021). Soluble Fas ligand drives autoantibody-induced arthritis by binding to DR5/TRAIL-R2. eLife, 10, e48840.

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